Kinectin 1 CRISPR Activation Plasmid (h): sc-404296-ACT

Human KTN1 encodes kinectin 1, an endoplasmic reticulum membrane–associated receptor for kinesin that helps couple microtubule-based transport to ER organization and dynamics. Through interactions with kinesin motors and cytoskeletal networks, kinectin contributes to vesicle trafficking, organelle positioning, and maintenance of ER morphology, processes that influence secretory capacity and intracellular signaling. KTN1-linked transport and ER homeostasis intersect with pathways governing protein folding and stress responses, including the unfolded protein response, and are relevant to phenotypes associated with disrupted neuronal connectivity and altered cellular architecture. Perturbation of ER–microtubule coupling is broadly implicated in neurobiology and cell migration, making KTN1 a useful node for mechanistic studies of intracellular transport regulation.

Kinectin 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous KTN1 expression without altering the underlying DNA sequence.

Kinectin 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the KTN1 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the KTN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Kinectin 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native KTN1 locus and enabling the study of Kinectin 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Kinectin 1 pathway restoration in tumor cells with silenced or reduced KTN1 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.